AAV phSyn1(S)-FlpO-bGHpA
(Plasmid #51669)

• Purpose: Can be used to generate AAV virus that will express the FlpO recombinase gene in neurons from the synapsin promoter.
• Depositing Lab: Hongkui Zeng
• Publication: Madisen et al Neuron. 2015 Mar 4;85(5):942-58. doi: 10.1016/j.neuron.2015.02.022.

Backbone

• Vector backbone: pAAV-MCS
• Backbone manufacturer: Stratagene
• Backbone size w/o insert (bp): 2887
• Total vector size (bp): 4972
• Modifications to backbone: A fragment containing the hSyn1 promoter-FlpO recombinase gene-bGHpA were swapped in to replace the CMV promoter-MCS-hGHpA sequence. Total insert size including the two ITRs is 2375 bp
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl2
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: FlpO recombinase gene
• Alt name: Flp recombinase gene codon optimized for expression in mice.
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 1299
• Promoter: hSyn1

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Not1 (not destroyed)
• 3′ cloning site: Not1 (not destroyed)
• 5′ sequencing primer: none
• 3′ sequencing primer: none

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Synthesized based on FlpO sequence in this reference: Raymond C.S. and P. Soriano.High-Efficiency FLP and ΦC31 Site-Specific Recombination in Mammalian Cells. 2007. PLoS One 2(1) e162.
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Information for AAV Retrograde (Catalog # 51669-AAVrg)

Purpose

Ready-to-use AAV Retrograde particles produced from AAV phSyn1(S)-FlpO-bGHpA (#51669). In addition to the viral particles, you will also receive purified AAV phSyn1(S)-FlpO-bGHpA plasmid DNA.

hSyn-driven FlpO expression. These AAV were produced with a retrograde serotype, which permits retrograde access to projection neurons. These AAV preparations are suitable purity for injection into animals.

Delivery

• Volume: 100 µL
• Titer: ≥ 7×10¹² vg/mL
• Pricing: $375 USD for preparation of 100 µL virus + $30 USD for plasmid.
• Storage: Store at -80℃. Thaw just before use and keep on ice.
• Shipment: Viral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.

Viral Production & Use

• Packaging Plasmids: encode adenoviral helper sequences and AAV rep gene, AAV retrograde cap gene from rAAV2-retro helper (plasmid #81070)
• Buffer: PBS + 0.001% Poloxamer 188 + 200 mM NaCl
• Serotype: AAV retrograde (AAVrg)
• Purification: Iodixanol gradient ultracentrifugation

Biosafety

Requestor is responsible for compliance with their institution's biosafety regulations. Lentivirus is generally considered BSL-2. AAV is generally considered BSL-1, but may require BSL-2 handling depending on the insert. Biosafety Guide

Terms and Licenses

• Academic/Nonprofit Terms:
  • Terms of Use for Viral Vectors
• Industry Terms:
  • Not Available to Industry

Viral Quality Control

Quality Control:

• Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). The specific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
• Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.

Visit our viral production page for more information.

Addgene Comments

Retrograde functionality is dependent on high viral titers. Addgene recommends not diluting your AAV preps prior to use.